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Biomedical subjects

K Park

Publications and source records attributed to K Park.

At least 343 records · Page 19Linked to original sources
PubMed1994

Idiopathic CD4+ T-lymphocytopenia and recurrent vulvar intraepithelial neoplasia.

BACKGROUND: CD4+ T-lymphocytopenia immunodeficiency without human immunodeficiency virus (HIV) infection has been reported recently. The association between immunodeficiency and anogenital neoplasia secondary to human papillomavirus infections is well documented. CASE: A woman with recurrent vulvar intraepithelial neoplasia (VIN) had idiopathic CD4+ T-lymphocytopenia without HIV infection. CONCLUSION: Human papillomavirus-related VIN may be associated with idiopathic CD4+ T-lymphocytopenia.

Carcinoma in Situ↗
PubMed1993

Characterization of the P4 promoter region of the human insulin-like growth factor II gene.

The human insulin-like growth factor II (IGF-II) gene contains four promoters (P1, P2, P3 and P4). In order to determine the mechanism by which the P4 promoter is controlled, the human IGF-II P4 promoter was analyzed in cell lines. DNA sequence analysis of the human IGF-II P4 promoter gene showed that the P4 promoter region contains a TATA-like sequence and several G+C rich regions which are essential for transcription. Analysis of the transcription initiation site by S1 nuclease mapping revealed two transcription start sites; both are located immediately behind TATA-like sequence. To determine the location of sites that may be important for the function of the human IGF-II P4 promoter, we constructed chimeric genes of the human IGF-II P4 promoter fused to the coding region for chloramphenicol acetyltransferase (CAT). These constructs were transfected into HepG2, PLC/PRF/5, G401 and A549 cells, and were examined for CAT activity. All transfected cells showed a similar profile of CAT activity. Sequences responsible for putative enhancer and silencer regions were identified and the 5' flanking sequences of the human IGF-II P4 promoter contain negative regulatory regions (-213 to -174). The 53-base pair fragment located between 111 and 59 base pairs upstream of the start site contains positive regulatory activity. Gel mobility shift assay showed that Sp1 and another proteins might be involved in positive regulation of the human IGF-II P4 promoter.

Animals↗
PubMed1993

Dynamic structural and functional relationships in recombinant plasminogen activator inhibitor-1 (rPAI-1).

The conformational characteristics of active, latent, and denatured recombinant plasminogen activator inhibitor-1 (rPAI-1) were compared using UV spectroscopy, spectrofluorimetry and circular dichroism (CD) techniques. The UV absorbance wavelength maxima in all preparations approximated 280 nm, while the extinction coefficients of active and latent rPAI-1 differed by up to 60%. When incubated at 37 degrees C, the A280 of latent rPAI-1 was quite stable while the A280 of active rPAI-1 spontaneously increased, eventually approximating that of latent rPAI-1. Alkali difference spectroscopy yielded markedly divergent titration patterns for active and latent rPAI-1, suggesting that the tyrosine residues present in active and latent rPAI-1 differ in terms of solvent exposure. At an excitation wavelength of 280 nm, active rPAI-1 exhibited the greatest relative fluorescence quantum yield. The relative fluorescence of latent and denatured rPAI-1 were less than that of active PAI-1, and the emission maxima of both species were slightly red-shifted in comparison to that of active rPAI-1, suggesting that at least one of the four tryptophan residues present in rPAI-1 is less exposed to the aqueous environment in the active form of the molecule. In contrast, the derived secondary structures based on CD of active and latent rPAI-1 were nearly identical, with both moieties exhibiting approx. 40% alpha-helix and 15% beta-sheet. Taken together, these spectroscopic data provide evidence supporting the hypothesis that active and latent PAI-1 differ in terms of their tertiary conformation and aromatic residue exposure, while their secondary structures appear generally comparable. Furthermore, denaturant-induced reactivation of latent rPAI-1 produces a partially active rPAI-1 with spectroscopic properties similar to that of latent rPAI-1, suggesting that denatured rPAI-1 more closely resembles the latent rPAI-1 conformation after refolding. The spontaneous spectroscopic changes observed in rPAI-1 may reflect conformational transitions that are critical to the regulation of endogenous PAI-1 activity.

Circular Dichroism↗
PubMed1993

The site of cAMP action in the insulin induction of gene expression of acetyl-CoA carboxylase is AP-2.

Insulin induction of acetyl-CoA carboxylase (ACC) and differentiation of 30A5 preadipocytes into adipocytes requires a brief exposure of the cells to cAMP. Using the techniques of DNase I footprinting, DNA band shift, and analysis of the point and deletion mutations, a region from -113 to -95 has been identified as the site through which cAMP sensitizes the cell for the response to insulin. One sequence-specific DNA-protein complex, b3, is formed in the DNA-mobility shift assay when nuclear extract from 30A5 cells is mixed with the oligonucleotide representing this region. Purified human AP-2 also generates the complex corresponding to b3 with the same ACC PII probe or with the AP-2 consensus sequence probe from SV40 promoter. Substitution of A for G in the sequence GGGGCTGGG abolishes the formation of b3 sequence-specific complex. Stably transfected 30A5 cells with the same mutations in the plasmid no longer respond to insulin in spite of their exposure to cAMP. These results establish that the 21 base pair region in ACC promoter II and the binding of AP-2 protein to this sequence are required for cAMP action. cAMP-dependent protein kinase phosphorylates AP-2 both in vitro and in vivo and the phosphorylation of AP-2 does not affect its binding activity.

Acetyl-CoA Carboxylase↗
PubMed1993

Identification of a transcriptional initiator element in the cytochrome c oxidase subunit Vb promoter which binds to transcription factors NF-E1 (YY-1, delta) and Sp1.

We have mapped the basal promoter activity of the mouse cytochrome c oxidase (COX) subunit Vb gene to the -17 to +20 region which contains two putative ets binding sites flanking an NF-E1 site fused to an Sp1 site. A 17-nucleotide sequence flanking the major transcription start site (-8 to +9), referred to as 17Inr (initiator sequence) was able to drive CAT activity in 3T3 cells to a level comparable to the construct containing sequences -17 to +20. This suggests that the 17Inr sequence contains the initiator activity. The 17Inr contains a pyrimidine-rich sequence, commencing with a CA that corresponds to the major transcription start site. Primer extension of RNA from transfected cells demonstrated that transcription initiation with the 17Inr template occurs at a site identical to the endogenous gene. DNA-protein binding by gel mobility shift and methylation interference analyses indicated that the pyrimidine-rich sequence immediately flanking the transcription start site consists of an NF-E1 factor binding motif with an overlapping upstream Sp1 binding site. A 13-nucleotide sequence, 13Inr (-4 to +9), which retains the NF-E1 binding activity but does not bind Sp1, was able to promote chloramphenicol acetyltransferase gene expression at levels similar to the 17Inr sequence, suggesting that NF-E1 factor binding is critical for initiator function. Finally, using an in vitro transcription system from Drosophila embryos we demonstrate that NF-E1 is necessary for transcription activation of both the 17Inr and the 13Inr initiator templates. Thus NF-E1 binding appears to be important for basal promoter function of the mouse COXVb gene.

3T3 Cells↗
PubMed1993

Conformational change of chaperone Hsc70 upon binding to a decapeptide: a circular dichroism study.

The conformation of bovine Hsc70, a 70-kDa heat shock cognate protein, and its conformational change upon binding to decapeptides, was studied by CD spectroscopy and secondary structure prediction (Chou, P.Y. & Fasman, G.D., 1974, Biochemistry 13, 222-245). The CD spectra were analyzed by the LINCOMB method, as well as by the convex constraint analysis (CCA) method (Perczel, A., Park, K., & Fasman, G.D., 1992, Anal. Biochem. 203, 83-93). The result of the CD analysis of Hsc70 (15% alpha-helix, 24% beta-sheet, 24% beta-turn, and 38% remainder) was very similar to the predicted secondary structure for the beta-sheet (24%) and the beta-turn (29%). However, there is disagreement between the alpha-helical content by CD analysis (15%) and the predicted structure (30%). In spite of the fact that the decapeptides contained a considerable amount of beta-sheet (22%), the interaction of the heat shock protein with the peptide resulted in an overall decrease in the content of beta-sheet conformation (-15%) of the complex. This may be due to induction of a molten globule state. The result of the CCA analysis indicated that the Hsc70 undergoes a conformational change upon binding the decapeptides.

Amino Acid Sequence↗
PubMed1993

Albumin grafting on to polypropylene by thermal activation.

4-azido-2-nitrophenyl albumin (ANP-albumin) was prepared by reacting 4-fluoro-3-nitrophenyl azide with albumin. The thermal decomposition kinetics of phenyl azide of ANP-albumin was studied at various temperatures by Fourier-Transform Infrared (FTIR) spectroscopy. The decomposition rate of the phenyl azide increased with temperature. The activation energy for the first-order decomposition of the phenyl azide was 128.0 kJ/mol. Albumin was grafted on to polypropylene (PP) films by thermolysis of the azido groups of ANP-albumin with no premodification of the PP surface. The albumin-grafted surface was characterized by electron spectroscopy for chemical analysis (ESCA) and by quantitative determination of platelet adhesion and activation. The bulk concentration of ANP-albumin used for adsorption varied from 0.001 to 30 mg ml-1, and the albumin-adsorbed PP films were incubated at 100 degrees C for up to 7 h. The carbon and nitrogen peaks resulting from the grafted albumin were used to compare the surface albumin concentrations as a function of the concentration of ANP-albumin in the adsorption solution. When the PP film was adsorbed with ANP-albumin at the concentration of 5 mg ml-1 or higher and incubated at 100 degrees C for longer than 5 h, the surface became resistant to platelet adhesion. The ANP-albumin can be grafted on to chemically inert surfaces such as PP surface through simple thermolysis of azido groups to prevent platelet adhesion and activation.

Adsorption↗
PubMed1993

The intracisternal A-particle upstream element interacts with transcription factor YY1 to activate transcription: pleiotropic effects of YY1 on distinct DNA promoter elements.

Murine intracisternal A-particle long terminal repeats contain an intracisternal A-particle upstream enhancer (IUE) element that binds to a 65-kDa IUE binding protein (IUEB) present in both undifferentiated F9 embryonal carcinoma cells and differentiated parietal endoderm-like PYS-2 cells. This IUE element confers a CpG methylation-sensitive IUEB binding and enhancer activity. Using gel retardation, methylation interference, CpG methylation sensitivity binding, and cotransfection assays, we have now identified the 65-kDa IUEB as YY1 (also called NF-E1, delta, or UCRBP), a zinc finger protein related to the Krüppel family. YY1 binds to a number of similar but distinct DNA motifs, and cotransfection assays indicate that these motifs have different enhancer potentials in PYS-2 cells. The relative strengths of these elements are as follows: IUE > kappa E3' from the human immunoglobulin kappa light-chain 3' enhancer > upstream conserved region from the Moloney murine leukemia virus promoter. Results of DNA binding assays suggest that the differences in enhancer potentials are due to the different binding affinities of YY1 to the various motifs and the binding of two other transcription factors to the IUE sequence.

Animals↗
PubMed1993

The relationship of monkey frontal eye field activity to saccade dynamics.

1. In this study, we compared the temporal waveforms of the activity of monkey frontal eye field movement neurons with the dynamics of saccadic eye movements. 2. Movement neurons in the frontal eye field were selected according to previously published criteria. They had little or no response to visual stimuli in a fixation task, and equivalent activity before visually guided and memory-guided saccades. We studied corticotectal neurons and corticopontine neurons identified by antidromic excitation, as well as neurons whose projections were not identified. 3. These neurons had a peak activation at a mean of 13 ms before the saccade began. However, rather than falling off rapidly as the saccade ended, most neurons continued to fire after the saccade, returning to baseline at a mean of 93 ms after the end of the saccade. 4. We measured the decrement in activity for these neurons during the saccade. Although a few neurons showed decrements of > 60% of their peak activity level, the average activity dropped only 16.9%, with some neurons actually showing a rise in activity during the saccade. If we ignored the latency between peak in activity and saccade start and measured the fall in activity for a period equal to one saccade duration after the peak, the average drop in activity was still only 34.9%. Thus the activity of these neurons did not appear to be closely related to dynamic motor error, which falls from its maximum value to zero over the time course of a saccade. 5. These results suggest that a focus of movement activity within the topographic map in the frontal eye field specifies the amplitude and direction for an impending saccade, whereas the peak of movement activity signals the time to initiate a saccade. 6. Unlike the superior colliculus, the activity of frontal eye field movement neurons does not appear to be related to dynamic events that occur during the saccade, such as motor error.

Animals↗
PubMed1993

Surface modification of polymeric biomaterials with poly(ethylene oxide), albumin, and heparin for reduced thrombogenicity.

Appropriate surface modification has significantly improved the blood compatibility of polymeric biomaterials. This article reviews methods of surface modification with water-soluble polymers, such as polyethylene oxide (PEO), albumin, and heparin. PEO is a synthetic, neutral, water-soluble polymer, while albumin and heparin are a natural globular protein and an anionic polysaccharide, respectively. When grafted onto the surface, all three macromolecules share a common feature to reduce thrombogenicity of biomaterials. The reduced thrombogenicity is due to the unique hydrodynamic properties of the grafted macromolecules. In aqueous medium, surface-bound water-soluble polymers are expected to be highly flexible and extend into the bulk solution. Biomaterials grafted with either PEO, albumin, or heparin are able to resist plasma protein adsorption and platelet adhesion predominantly by a steric repulsion mechanism.

Albumins↗
PubMed1993

Development of secretory elements in murine tubotympanum: lysozyme and lactoferrin immunohistochemistry.

Secretory activity, particularly of antibacterial agents such as lysozyme and lactoferrin, is an important aspect of the mucosal defense mechanism, and the development of these agents may have a direct bearing on the susceptibility of the ear to infection. In this study, the secretory cells of the murine tubal epithelium were first observed at gestational day 17. Although tubal glands began to develop on gestational day 18, their secretory activity was first shown on postnatal day 3. The number of secretory cells of the tubal epithelium increased rapidly immediately after birth, while that of the tubal glands showed a gradual increase from postnatal day 3. The epithelial secretory cells in the tubotympanum matured at birth, but the tubal glands matured gradually after birth. Lysozyme was first recognized in the epithelial secretory cells on postnatal day 1, while lactoferrin was first detected in the tubal glands on postnatal day 3. Both lysozyme and lactoferrin were co-localized in the serous cells of the tubal glands. The secretion of lysozyme and lactoferrin seemed to reflect the maturation of the secretory cells in the murine tubotympanum.

Alcian Blue↗
PubMed1993

Effect of influenza A virus on ciliary activity and dye transport function in the chinchilla eustachian tube.

Previous studies have shown that influenza A virus inoculated via a transbullar approach can cause severe disorders of the chinchilla eustachian tube. In this study, we inoculated chinchillas both intranasally and transbullarly with influenza A virus (A/Alaska/6/77) to assess the effect of this virus on both the ciliary activity of eustachian tube epithelium and eustachian tube transport function. Data indicated that while the nature of the major lesions was indistinguishable, the differing routes of inoculation did influence the time course and initial location of virus-induced histopathology. Analysis of ciliary beat frequency and dye transport function indicated a maximal decrease approximately 7 to 14 days postinoculation, with return to normal function by 28 days postinoculation. Light and transmission electron microscopic observations of the evolution of histopathology and return to normal histology correlated well with functional assessment data.

Administration, Intranasal↗
PubMed1993

Photografting of albumin onto dimethyldichlorosilane-coated glass.

4-Azido-2-nitrophenyl albumin (ANP-albumin) was prepared by displacing the fluoro group of 4-fluoro-3-nitrophenyl azide (FNPA) by an amino group of albumin. Photolysis of phenyl azides of ANP-albumin was studied by Fourier-transform infrared (FTIR) spectroscopy. The band of phenyl azide disappeared completely after a 12-min exposure to long wave UV light (366 nm), and the photolysis was first-order. Albumin was grafted onto dimethyldichlorosilane-coated glass (DDS-glass) by photolysis of the azido groups of ANP-albumin without any premodification of the surface. The albumin-grafted DDS-glass was characterized by determining the relative amount of nitrogen resulting from the grafted albumin on the surface using electron spectroscopy for chemical analysis (ESCA). The amount of nitrogen increased when the concentration of ANP-albumin in the adsorption solution increased up to 0.1 mg/ml. As the solution concentration increased above this value, the amount of nitrogen decreased. The platelet resistance of the albumin-grafted surfaces was evaluated by measuring the number of adherent platelets and the extent of activation that was quantitated by the area of platelets spread on the surfaces. The maximum platelet-resistant effect was observed when the ANP-albumin was adsorbed for more than 50 min at the solution concentration ranging from 0.05 to 10 mg/ml.

Adsorption↗
PubMed1993

An element of the transforming growth factor-beta 1 5'-untranslated region represses translation and specifically binds a cytosolic factor.

In many cell types, there is a discrepancy between transforming growth factor-beta 1 (TGF-beta 1) mRNA and TGF-beta 1 protein, suggesting that expression of TGF-beta 1 is regulated posttranscriptionally. We have previously shown that a 137-nucleotide (nt) region of the TGF-beta 1 5'-untranslated region (UTR) potently inhibits the expression of a heterologous reporter gene, suggesting a role for this region in the posttranscriptional inhibition of TGF-beta 1 expression. To study the mechanism of inhibition, a chimeric plasmid containing this region of the TGF-beta 1 5'-UTR and the reading frame of the human GH gene was stably transfected into C2C12 myoblastic cells. Our results show that the TGF-beta 1 5'-UTR inhibits GH expression by inhibiting GH mRNA translation. In vitro gel retardation and cross-linking assays using a radiolabelled RNA probe transcribed from this region of the TGF-beta 1 5'-UTR demonstrate the specific binding of a cytosolic factor. Deletion of a potential stem-loop-forming region abolishes binding of this factor and partially restores GH production. These results suggest that posttranscriptional inhibition of TGF-beta 1 expression is at the level of mRNA translation and that a cytosolic factor may regulate TGF-beta 1 mRNA translation.

Animals↗
PubMed1993

Living donor nephrectomies-right side--intraoperative assessment of the right renal vascular pedicle in 112 cases.

Generally, the left kidney from a living donor is more commonly preferred but the right kidney is occasionally donated because of multiple left renal arteries or repeated transplantation. The right donor nephrectomy is usually more difficult compared to the left because the right renal vein is often multiple and short, which complicates dissection of the vascular pedicle. From Jan. 1989 to Sep. 1992, 112 consecutive cases of right donor nephrectomies out of a total of 771 cases were retrospectively reviewed with the preoperative renal angiography and the intraoperative assessment of the right renal vascular pedicle. The indications for right donor nephrectomy include 1) multiple or proximal bifurcating renal arteries of the left kidney (89.3%), 2) repeated transplantation in the recipient (9%). In 26.8% of the cases, there were more than two right renal veins. In the right donor nephrectomy, it is often necessary to perform vena cava cuff resection because of short and frequently occurring multiple right renal veins. For the dissection of the inferior vena cava (IVC), the aberrantly occurring right gonadal vein, the adrenal vein draining above the junction of the renal vein and IVC, and the lumbar vein below the junction should always be looked for and must be ligated if any are found.

Adult↗